TABLE 1

Biochemical features of the wild type and the evolved UPO variant

Biochemical or spectroscopy featurehValue
wtUPO1ePaDa-I mutantf
Mass (Da)a46,00052,000
Mass (Da)bNDg51,100
Mass (Da)c35,94235,914
Glycosylation degree (%)2230
Thermal stability (T50) (°C)d5355
pI4.9–5.75.5
Optimum pH for ABTS4.04.0
Optimum pH for DMP7.06.0
Optimum pH for NBD6.56.0
Rz (A418/A280)2.41.8
Soret region (nm)420418
CT1 (nm)572570
CT2 (nm)540537
  • a Estimated by SDS-PAGE.

  • b Estimated by MALDI-TOF mass spectrometry.

  • c Estimated from the amino acid composition.

  • d Estimated from purified variants.

  • e wtUPO1, UPO1 wild type expressed in A. aegerita.

  • f PaDa-I mutant, the ultimate variant of the whole evolution process in S. cerevisiae (containing the evolved signal peptide [n*] plus the evolved UPO1) (see Fig. S1 in the supplemental material).

  • g ND, not determined.

  • h CT1 and CT2, charge transference bands 1 and 2, respectively.